[丙型肝炎中的聚合酶链反应：通过扩增较短的cDNA片段提高敏感性]

[Polymerase chain reaction in hepatitis C: increased sensitivity via amplification of shorter cDNA fragments].

作者信息

Schwery S, Wicki A N, Joller-Jemelka H I, Havelka J, Zala G, Schmid M, Bühler H

机构信息

Medizinische Klinik, Stadtspital Waid, Zürich.

出版信息

Schweiz Med Wochenschr. 1993 May 15;123(19):965-9.

PMID:8511526

Abstract

The aim of our study was to compare the sensitivity of hepatitis C virus polymerase chain reaction (HCV-PCR) by use of two different primer sets which amplify PCR products of different length. Serum samples of 70 patients with chronic hepatitis C were tested by "nested primer" PCR, using either "NCR primers" that amplify cDNA-fragments of 340 basepairs (bp), or by "PT primers" which amplify fragments of 59 bp only. HCV-RNA was detected in 40 patients (57%) by "NCR primers" and in 69 patients (90%) by "PT primers" (p < 0.001). 23 of 70 patients (33%), which were HCV-RNA negative by "NCR primers", were positive by "PT primers", but no patient negative by "PT primers" was found to be positive by "NCR primers". 20 healthy controls tested by both primer sets were all HCV-RNA negative. We conclude that the sensitivity of HCV-PCR is significantly improved by use of primers that amplify "short" PCR products and recommend the use of "PT primers" for HCV-PCR.

摘要

我们研究的目的是比较使用两种不同引物组的丙型肝炎病毒聚合酶链反应（HCV-PCR）的敏感性，这两种引物组扩增的PCR产物长度不同。对70例慢性丙型肝炎患者的血清样本进行“巢式引物”PCR检测，使用扩增340个碱基对（bp）cDNA片段的“NCR引物”，或仅扩增59 bp片段的“PT引物”。通过“NCR引物”在40例患者（57%）中检测到HCV-RNA，通过“PT引物”在69例患者（90%）中检测到HCV-RNA（p<0.001）。70例患者中有23例（33%）通过“NCR引物”检测HCV-RNA为阴性，但通过“PT引物”检测为阳性，然而未发现通过“PT引物”检测为阴性的患者通过“NCR引物”检测为阳性。对20名健康对照使用两种引物组进行检测，结果均为HCV-RNA阴性。我们得出结论，使用扩增“短”PCR产物的引物可显著提高HCV-PCR的敏感性，并建议在HCV-PCR中使用“PT引物”。